| The classical formula Yiguanjian Decoction is a famous formula created by Zhixiu Wei in the Qing Dynasty for the treatment of liver diseases,which is still widely used in clinical practice and has great development value due to its precise efficacy.2018,the state announced the first batch of 100 formulas derived from ancient classical prescriptions,and Yiguanjian Decoction was among them.Based on the high-quality requirements for the development of classical prescriptions,the state issued a series of guidelines to regulate the development of classical prescriptions,and also pointed out that quality control research is one of the technical difficulties in the development of classical prescriptions.Quality control is an important tool to ensure the quality of classical prescriptions throughout the whole process of their development.However,due to the complexity of classical prescriptions,the quantitative control can only select the components with high content for quantitative control,which is difficult to reflect the quality traceability of all herbs.Therefore,this topic proposes a quantitative control research idea based on the full coverage of the whole process,through the material basis of the Yiguanjian decoction system characterization,screening the relatively high content of the characteristic components of the Yiguanjian decoction representing each herb,selecting the current Chinese Pharmacopoeia acceptable high performance liquid chromatography-single quadrupole mass spectrometry to establish a quantitative control method,in order to explore the full coverage of the quantitative control idea for the quality evaluation of the whole process of monitoring the production of classic prescriptions.In order to explore the role of full-coverage quantitative control ideas for the quality evaluation of the whole process of the production of classic prescriptions,and provide new ideas and methods for the improvement of the quality of classic prescriptions.This research is divided into the following four aspects:1.Comprehensive characterization of the chemical composition of Yiguanjian Decoction.Based on ultra performance liquid chromatography tandem linear ion trap electrostatic field orbital high resolution mass spectrometry(UHPLC/LTQ-Orbitrap HRMS)technology,the high resolution mass spectrometry data of Yiguanjian decoction,single herbs and their negative preparations were firstly collected,and then the rapid matching method was achieved through the identification of controls,literature reports,and high resolution liquid-quality data processing software(MSDIAL,V4.92).The results showed that the aqueous extracts of Shengdi and its monosodium glutamate were fully characterized.The results showed that 131 compounds were identified in the aqueous extracts of Radix et Rhizoma,including cyclic enol ether terpene glycosides,phenylethanol glycosides,sugars and violet ketones;116 compounds were identified in the aqueous extracts of Lycium barbarum,including spermidine,phenylpropanoids,organic acids and amides;95 compounds were identified in the aqueous extracts of Angelica sinensis,including phthalides,organic acids,amino acids,sugars and nucleosides;95 compounds were identified in the aqueous extracts of Angelica sinensis,including phthalides,organic acids,amino acids,sugars and nucleosides.A total of 88 compounds,mainly steroidal saponins and high isoflavones,were identified in the aqueous extract of Radix Angelicae Sinensis;28 compounds,mainly coumarins and organic acids,were identified in the aqueous extract of Radix Salviae Sinensis;34 compounds,mainly triterpenoids,were identified in the aqueous extract of Neem.A total of 388 compounds were identified in a Yiguanjian Decoction,including 123 compounds from Shengdi,86 from Lycium,65 from Medlar,64 from Angelica,29 from Neem,and 21 from Bei Sha Shen;including 55 cyclic enol ether terpenes,43 spermidine,42 steroidal saponins,17 triterpenoids,29 phenylpropanoids,28 organic acids,18 violet ketones,and high 15 isoflavones,15 coumarins,14 phenylethanol glycosides,14 sugars,9 amino acids,8flavonoids,8 alkaloids,and 52 other components.2.Selection and determination of high-coverage quantitative characteristic components of Yiguanjian Decoction.To address the difficulties in the selection of high-coverage index components in the quality control study of Yiguanjian Decoction,an effective strategy and method for the selection of high-coverage index components of Yiguanjian Decoction was constructed based on the high-resolution liquid mass spectrometry data processing software(Progenesis QI,V2.4).The method takes the abundance value(Abundance)in the derived peak table of Progenesis QI software as the calculation index,and carries out the conditions of(1)AYGJ-ABlank ≥ 5104(to remove background interference and to ensure high abundance),(2)Asingle-drug-AYGJ≥0(to ensure strong features),(3)AYGJ/Anegative ≥ 102(negative comparison,to ensure specificity)in order to Feature ion screening,combined with mass loss filtering(MDF)technology and full-component characterization basis,a total of 13 high-coverage quantitative and semi-quantitative feature components were identified,covering 7types of compounds,and all medicinal flavors were monitored to achieve the goal of high-coverage quality control.3.The establishment of a Yiguanjian Decoction high coverage quality control method.Based on ultra performance liquid chromatography tandem with a compact single quadrupole mass spectrometer(UPLC/QDa),this technique has the advantages of high sensitivity,easy operation,low cost,high practicality and wide analytical range.A high coverage quantitative analysis method was established by applying Multiple Ion Monitoring(MIM)mode to simultaneously monitor 13 characteristic components in a Yiguanjian Decoction.The method was validated,and the linearity of the 8 quantitative components was good in the set concentration range,and the RSD values of repeatability,stability and inter-day precision were less than 8%.The average recoveries of the samples ranged from 79.88% to 116.10%,which were in accordance with the requirements of Chinese Pharmacopoeia.4.Whole-process sample quality analysis of Yiguanjian Decoction.The established quantitative analysis method of UPLC/QDa was used to analyze the quality of the decoction in the whole production process.The results showed that firstly,the established quantitative analysis method could stably detect 13 index components in the samples of each process and achieve the goal of monitoring the quality of samples in the whole production process of Yiguanjian Decoction.Secondly,for the first time,the effects of the whole process of Yiguanjian decoction production(herbs/drink extracts,intermediates and preparations)on the full coverage of quantitative indexes were found,such as the Ligustilide in Angelica sinensis was influenced by the drying process,and the pepper toxin was influenced by the skin layer of the herbs.This research method better demonstrates that different herbs are influenced by different processes and shows better quality control accuracy compared with traditional quality control.The above study shows that the quality control strategy based on full coverage and full process quantification can reflect the changes of all medicinal flavors in the production process of classical formulae,can better trace each herb in the final product quantitatively,and can understand the consistency of the product more precisely,which shows the unique advantages of this research strategy and provides a new idea and methodological reference for the quality control research of complex system of classical formulae. |
PDF Full Text Request